CN106442483A - Luminous bacterium flow injection method for quickly detecting and warning food-borne toxin pollution and application of luminous bacterium flow injection method - Google Patents
Luminous bacterium flow injection method for quickly detecting and warning food-borne toxin pollution and application of luminous bacterium flow injection method Download PDFInfo
- Publication number
- CN106442483A CN106442483A CN201611041464.2A CN201611041464A CN106442483A CN 106442483 A CN106442483 A CN 106442483A CN 201611041464 A CN201611041464 A CN 201611041464A CN 106442483 A CN106442483 A CN 106442483A
- Authority
- CN
- China
- Prior art keywords
- flow injection
- luminous
- detection
- food
- borne
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
- G01N21/763—Bioluminescence
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
- G01N35/08—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor using a stream of discrete samples flowing along a tube system, e.g. flow injection analysis
- G01N35/085—Flow Injection Analysis
Abstract
The invention belongs to the field of food safety, environment monitoring and public safety, and particularly discloses a luminous bacterium flow injection method for quickly detecting and warning food-borne toxin pollution and application of the luminous bacterium flow injection method. The luminous bacterium flow injection method includes steps of resuscitating, inoculating and continuously cultivating vibrio qinghaiensis Q67 microbial agents; designing flow injection system programs; detecting sample solution by the aid of luminous bacterium flow injection processes; integrating and processing data and designing warning systems. The luminous bacterium flow injection method and the application have the advantages that the food-borne toxin pollution can be detected and warned by the aid of the created luminous bacterium flow injection method, the luminous bacterium flow injection method is high in analysis speed and automation degree, easy and convenient to implement and good in repeatability, and errors due to manual operation can be prevented; flow injection analysis processes are combined with luminous bacterium continuous cultivation, and accordingly the possibility of cross pollution is extremely low; vibrio qinghaiensis is used as an indicator for measuring pollution toxin and is wide in detection range and sensitive in reaction.
Description
Technical field
The invention belongs to food safety and environmental monitoring and public safety field, and in particular to a kind of quick detection and early warning
The flow injection Luminous bacteria of food-borne endotoxin contamination and application.
Background technology
Occur in recent years sudden quote in water contamination accident, artificial drinking water is poisoned and is difficult to be found in time, some
Common food-borne toxin strong toxicity and simple and easy to get, it is easy to utilized by lawless person, hence set up food source in drinking water
Property toxin quick detection and early warning system are significant.
Chemical analysis can carry out quantification and qualification to pollutant in water environment, but can not directly reflect pollutant pair
Environment and biological impact.Toxotest method with photobacteria as biological subject, because quick, easy, sensitive, stable, economical
And many experiments that receive publicity show, photobacteria toxicity test is tested to having in toxicity of compound evaluation well with toxicity in fish
Dependency.Luminescent bacteria is sensitive to the toxic reaction in environment, and after showing contact poison, luminous intensity can be suppressed rapidly.
There are following characteristics in China distinctive luminescent bacterium Vibrio-qinghaiensis sp. Q67 (Vibrioqinghaiensis sp.Q67)：(1) it is not
Needing NaCl to exist also can be good luminous (can carry out in fresh water system normal growth and light)；(2) adaptive temperature scope
Extensively；(3) pH value in interior equal energy growth in a big way and lights；(4) show it for non-pathogenic bacteria to the infection experiment of Carassius auratuses.For
The monitoring Qinghai Vibrion of general poisons in freshwater or pollutant has the advantages which is unique.In recent years Qinghai Vibrion is received significant attention, but
The method for matching therewith still disunity, how using the luminous rear direct detection water quality toxicity of antibacterial lyophilized powder recovery.And adopt antibacterial
There is problems with lyophilized powder recovery approach：One is that lyophilized powder cost of manufacture is higher；Two is that anabiosis rate difference can affect detection knot
Really；Three is to carry at normal temperatures.Above feature limits its popularization and application.
Content of the invention
In place of solving the shortcoming and defect of prior art, the primary and foremost purpose of the present invention be to provide a kind of quick detection and
The flow injection Luminous bacteria of early warning food-borne endotoxin contamination.
Another object of the present invention is to the flow injection for providing above-mentioned quick detection and early warning food-borne endotoxin contamination is sent out
The application of photobacteria method.
The object of the invention is achieved through the following technical solutions：
A kind of quick detection and the flow injection Luminous bacteria of early warning food-borne endotoxin contamination, comprise the steps：
(1) recovery of Vibrio-qinghaiensis sp. Q67 microbial inoculum, inoculation and continuous culture；
(2) flow injection system programming；
(3) flow injection Luminous bacteria detection sample solution；
(4) data integration and process and the design of warning system.
Preferably, the concrete steps of the flow injection Luminous bacteria of the quick detection and early warning food-borne endotoxin contamination
As follows：
(1) recovery of Vibrio-qinghaiensis sp. Q67 microbial inoculum, inoculation and continuous culture：Take -20 DEG C of Vibrio-qinghaiensis sp. Q67 lyophilized powder ampoules 1
, after sterilization, 4 DEG C of pre-cooling 15min, resuscitation fluid slight oscillatory is added, 1000 μ L bacteria suspensions is inoculated with superclean bench extremely
In 1000mL fluid medium, 25 DEG C, 120rpm shaken cultivation 16h to optimal luminescent state, obtain testing with bacterium solution and shifting
Continuously cultivated to continuous cultivation and fermentation tank；
(2) flow injection system programming：Flow injection injector program setting such as table 1；
Table 1 sets flow injection injector program
(3) flow injection Luminous bacteria detection：The test bacterium solution that step (1) is continuously cultivated is entered by flow injection
Sample device pumps into superweak luminescence detecting system, using ultra-pure water as blank, after obtaining stable luminous signal, imports to be measured
Sample solution carries out luminous detection and obtains corresponding luminous intensity；
(4) data integration and process and the design of warning system：Data integration and process and warning system are by being sequentially connected
Photomultiplier tube, opto-electronic conversion and voltage amplification module, analog-to-digital conversion module, detection by quantitative module and alarm composition；Step
Suddenly after the optical signal that (3) detection is obtained amplifies through photomultiplier tube, by opto-electronic conversion and voltage amplification module, then through mould
Number modular converter obtains digital signal, obtains relative luminous intensity through detection by quantitative module；Relative luminous intensity=(sample lights
Intensity/control luminous intensity) × 100%, the triggering when detection by quantitative module detects relative luminous intensity less than 80% is reported to the police
Device is reported to the police.
Preferably, step (1) fluid medium is prepared according to the following steps and is transferred to feed supplement after carrying out sterilization treatment
In fluid reservoir：Weigh tryptone 5g, yeast powder 5g, NaCl 6.73g, Na2CO30.99g、NaHCO30.57g、MgCl2·
6H2O 2.92g、MgSO42.47g、CaCO30.03g, NaBr 0.1g and KCl 0.22g, adds the triangle of 1L to burn one by one
In bottle, glycerol adding 3mL, plus distilled water 1000mL, elimination precipitation was mixed, with the rear 121 DEG C of sterilizings 15min of kraft paper wrapping.
Preferably, step (3) sample solution is the aqueous solution containing 3-nitropropionic acid or containing Colchicine.
Preferably, step (3) sample solution be drinking water, the water source of food processing enterprises, Caulis Sacchari sinensis sample solution or
New hemerocallis sample solution.
Preferably, when step (3) sample solution is drinking water, in step (4), relative luminous intensity is below 80%
Then show that drinking water is potentially contaminated, alarm equipment alarm is triggered after detection by quantitative module analysis.
It is furthermore preferred that the Caulis Sacchari sinensis sample solution or new hemerocallis sample solution can be obtained by following steps：Weigh
After 10g Caulis Sacchari sinensis or new hemerocallis carry out pre-treatment, by supersound extraction 15min, quantitative filter paper is filtered, and filtrate is through 5000rpm
Standby with 0.2 μm of hole membrane filtration after high speed centrifugation.
It is furthermore preferred that the Caulis Sacchari sinensis pre-treatment refers to for extruding after testing sample peeling chopping to obtain sample liquid；Described new
Hemerocallis pre-treatment refers to be crushed with homogenizing instrument.
Preferably, in step (3) detection process, test bacterium solution and sample solution need premixing in advance then by
Phosphate buffer is loaded into superweak luminescence detecting system and is detected.
It is furthermore preferred that step (3) sample solution incorporation time in detection process is 10s with test bacterium solution, inspection
The survey time is 60s.Detection time be highly shortened compared to other detection methods, it is adaptable to the quick inspection of burst Pollution
Survey.
Preferably, in step (3) detection process, test bacterium solution feed rate is according to flow injection system detection speed
Degree adjustment remains the luminous intensity of relative constancy to ensure bacterium solution.
The flow injection Luminous bacteria of above-mentioned quick detection and early warning food-borne endotoxin contamination can be used for food processing enterprise
Sudden food-borne endotoxin contamination detection in the pollution of waterhead monitoring of industry, drinking water on-line monitoring and drinking water.
Compared with prior art, the present invention has advantages below and beneficial effect：
(1) chemical analysis can carry out quantification and qualification to pollutant in water environment, but can not directly reflect pollution
Thing is to environment and biological impact.Mostly thin for the detection method of food-borne toxin such as 3-nitropropionic acid, Colchicine at present
Layer chromatography, high performance capillary electrophoresis, gas chromatography, high performance liquid chromatography, Solid-Phase Extraction-Ultra Performance Liquid Chromatography series connection matter
Spectrometry etc..These methods exist and need to use the big organic reagent of a large amount of toxicity, complex operation, the shortcomings of detection cycle is long, only
Test in laboratory is can apply to, and be may not apply in productive life.Compare with said method, flow injection of the present invention is luminous thin
Bacterium method is due to simple to operate, sensitivity height, and can realize continuous sample introduction carries out on-line monitoring, can be widely used in food processing, ring
The field such as border monitoring and public safety.
(2) present invention is set up the detection of flow injection Luminous bacteria and early warning food-borne endotoxin contamination, analyze speed
Hurry up, easy to operate, high degree of automation, it is to avoid the error that manual operations causes, favorable reproducibility, due to being divided using flow injection
Continuously culture combines for analysis method and photobacteria, and the cross-contamination probability being subject to is very low.Meanwhile, by the use of Qinghai Vibrion as survey
The indicator of toxin is polluted surely, and detection range is wide, be quick on the draw.
Therefore, it can according to different detection demands, the contamination monitoring of environment water resource is applied to, sudden drinking water
Food-borne endotoxin contamination, or with the pollution of waterhead monitoring by optimizing and being modified to each production link of food processing enterprises.
Such continuous on-line monitoring system, can not only adjust the program of flow injection system and continuous culture dress according to actual needs
The monitoring for controlling to realize real-time detection and emphasis period and link that puts, and selection can have been carried out to specific pollutant
Property emphasis monitoring, monitoring for enterprise's production link and the early warning monitoring of sudden drinking water pollution have widely should very much
With prospect and practical value；Can to drinking water, sudden food-borne endotoxin contamination carries out early warning and sound on a large scale in the very first time
, can should effectively prevent such public safety hazards.
(3) direct detection water quality toxicity after typically being lighted using the recovery of antibacterial lyophilized powder at present.And adopt antibacterial lyophilized powder
There is problems with recovery approach：One is that lyophilized powder cost of manufacture is higher；Two is that anabiosis rate difference can affect testing result；Three are
Can not carry at normal temperatures.Above feature limits its popularization and application.
The present invention sets about from luminescent bacterium growth luminescence law, using the luminescent bacteria of fresh continuous culture, in conjunction with flowing
Injection luminescence method is studied to food-borne acute toxicity in water, is carried out using the luminous good fresh bacterium solution of logarithmic (log) phase growth
Experiment detection efficiently solves lyophilized powder cost of manufacture height, anabiosis rate difference to experiment and the restriction of Technique Popularizing, to for drinking
Detection foundation is provided with water food-borne toxin burst Pollution.
(4) Flow Injection Analysis (FIA) are under thermodynamics non-equilibrium condition, drive reproducing technology by current-carrying to process
The dynamic analysis technology of sample or reagent zone, with analyze speed is fast, accuracy and precision is good, high degree of automation
And the advantages of can combine with multiple detection meanss.The method of flow injection is combined by the present invention with Luminous bacteria,
The toxin in quick on-line monitoring drinking water can be realized；And flow injection photobacteria detection method, photobacteria and pollution toxin
Time of contact the time-limited problem of the short stabilized illumination for effectively overcoming Vibrio-qinghaiensis sp. Q67 bacterial strain, as a result more science is credible.
Description of the drawings
The step of Fig. 1 is flow injection photobacteria quick detection and early warning food-borne toxin method flow chart.
Fig. 2 is the installation drawing of the flow injection Luminous bacteria of quick detection and early warning food-borne endotoxin contamination, wherein：1-
Feed supplement fluid reservoir, 2- air filter, 3- air induction conduit, 4- peristaltic pump, 5- cultivation and fermentation tank, 6- water-bath constant temperature oscillator,
8- buffering liquid groove, 7- sample solution groove, 9- peristaltic pump, 10- mixing tube, 11- six-way valve, 12- waste liquid pool, 13- flow cell, 14-
Negative high voltage, 15- photomultiplier tube, 16- computer, 17- alarm.
The impact that Fig. 3 is grown to photobacteria for fluid medium pH.
The impact that Fig. 4 is grown to photobacteria for fluid medium temperature.
Fig. 5 is impact of the variable concentrations 3-nitropropionic acid to photobacteria relative luminous intensity.
Fig. 6 is impact of the variable concentrations Colchicine to photobacteria relative luminous intensity.
Fig. 7 is data integration and process and warning system schematic diagram.
Specific embodiment
With reference to embodiment and accompanying drawing, the present invention is described in further detail, but embodiments of the present invention are not limited
In this.
The flow injection Luminous bacteria of a kind of quick detection of the present invention and early warning food-borne endotoxin contamination includes following step
Suddenly：The recovery of Vibrio-qinghaiensis sp. Q67 microbial inoculum, inoculation and continuous culture；Flow injection system programming；Flow injection Luminous bacteria
Detection sample solution；Data integration and process and the design of warning system.The Vibrio-qinghaiensis sp. Q67 liquid for preparing is trained by the method
Foster base pumps into thalline fermentation culture tank through feed supplement fluid reservoir, and (as Qinghai Vibrion is aerobic bacteria, whole culture of continuous cultivation has
Filtrated air is pumped into), the size of food-borne endotoxin contamination and toxicity is judged whether through relative luminous intensity.
For realizing the device of the flow injection Luminous bacteria of quick detection of the present invention and early warning food-borne endotoxin contamination
As shown in Fig. 2 it includes photobacteria continuous culture system, sample adding system, superweak luminescence detecting system, data integration and place
Reason and warning system；Photobacteria continuous culture system is by feed supplement fluid reservoir 1, air filter 2, peristaltic pump 4, air induction conduit
3rd, cultivation and fermentation tank 5 and water-bath constant temperature oscillator 6 constitute, air induction conduit 3, air filter 2, feed supplement fluid reservoir 1, peristaltic pump
4th, cultivation and fermentation tank 5 is sequentially connected logical, and cultivation and fermentation tank 5 is placed in water-bath constant temperature oscillator 6；Sample adding system enters for flow injection
Sample device, including sample solution groove 7, peristaltic pump 9, buffering liquid groove 8, mixing tube 10, six-way valve 11, waste liquid pool 12 and flow cell 13,
Sample solution groove 7, buffering liquid groove 8, cultivation and fermentation tank 5 are connected with mixing tube 10 respectively, control test to use bacterium by peristaltic pump 9
Liquid, buffer, the flow acceleration of sample solution, control the mixing of bacteria liquid sample solution on probation and buffer by six-way valve 11
Sequentially, mixing tube 10, waste liquid pool 12 are connected with flow cell 13；Superweak luminescence detecting system include superweak luminescence detector and
Negative high voltage 14；Cultivation and fermentation tank 5 is connected with flow injection injector by flexible pipe；Data integration and process and warning system by
Photomultiplier tube 15, computer 16 and alarm 17 constitute.Computer 16 refers to that opto-electronic conversion and voltage amplification module, modulus turn
It is the detection program of computer settings that mold changing block and detection by quantitative module are arranged, after digital signal is obtained, through being compiled
The computer program that writes is carrying out detection by quantitative.
Following examples are application of the flow injection Luminous bacteria of the present invention in several food-borne toxin sample are detected.
Vibrio-qinghaiensis sp. Q67 lyophilized powder used in embodiment is purchased from shore pine photonic propulsion commerce and trade (China) company limited；Superweak luminescence is examined
Survey BPCL-K type Weak light investigating instrument of the instrument using Beijing Ya Bo Si Science and Technology Ltd.；Flow injection injector adopts Xi'an
The IFIS c-type intelligence flowing sample injection device of Rui Mai analytical tool Co., Ltd, adjusts current-carrying liquid stream with programme-control
Speed and the function of micro-sampling.
Embodiment 1
Food-borne toxin 3-nitropropionic acid, the detection example of Colchicine, concrete steps flow process is as shown in figure 1, including bacterium
The recovery of agent and the preparation of culture medium, test the preparation of bacterium solution, the preparation of toxin sample, luminous detection, the meter of relative method intensity
Calculate and analyze, the response of early warning system.Comprise the following steps that：
(1) preparing Toxin stock liquid and appropriate dilution is carried out to storing solution obtains required toxin sample concentration：
3-nitropropionic acid storing solution (500mg/L)：0.05g 3-nitropropionic acid (Sigma company) is weighed, and minister is bent with a small amount of
Family name distills water dissolution, is settled in 100mL volumetric flask and stores for future use.According to experiment need to dilute storing solution respectively obtain 100,
80th, 60,40,20, the 3-nitropropionic acid solution for standby of 10mg/L.
Colchicine storing solution (500mg/L)：0.05g Colchicine (Sigma company) is weighed, and dilution is needed according to experiment
Storing solution respectively obtains 10,8,6,4,2, the colchicine solution of 1mg/L standby.
(2) Caulis Sacchari sinensis sample solution and new hemerocallis sample solution are prepared：
Rubescent Caulis Sacchari sinensis sample and each 10g of new hemerocallis sample that after weighing peeling, chopping extruding is obtained, it is fixed to add water respectively
Hold to 100mL.Supersound extraction 15min, quantitative filter paper is filtered, and takes filtrate with 0.2 μm of microporous filter membrane after 5000rpm high speed centrifugation
Filter standby.Gained Caulis Sacchari sinensis sample solution or new hemerocallis sample solution and standard solution (i.e. 3-nitropropionic acid solution, autumn waters -- limid eyes
Celestial aqueous slkali) determine under identical conditions.
(3) Vibrio-qinghaiensis sp. Q67 fluid medium is prepared：
Tryptone 5g, yeast powder 5g, NaCl 6.73g, Na are weighed with electronic balance2CO30.99g、NaHCO3
0.57g、MgCl2·6H2O 2.92g、MgSO42.47g、CaCO30.03g, NaBr 0.1g and KCl 0.22g, one by one plus
Enter in the conical flask of 1L, glycerol adding 3mL, plus distilled water 1000mL, elimination precipitation was mixed, was wrapped up with kraft paper latter 121 DEG C
Sterilizing 15min.After cooling using or to be positioned over 4 DEG C of Refrigerator stores standby.
(4) recovery of Vibrio-qinghaiensis sp. Q67 microbial inoculum and inoculation, continuous culture：
4 DEG C of Vibrio-qinghaiensis sp. Q67 lyophilized powder ampoules 1 are taken, after cotton ball soaked in alcohol wipes periphery sterilization, emery wheel in superclean bench
Osculum is cut, ampoule is slowly opened, the 0.9%NaCl 1mL of 4 DEG C of pre-cooling 15min is added in the ampoule that opens, slight oscillatory, in
1000 μ L bacteria suspensions of inoculation are to 1000mL fluid medium in superclean bench, 25 DEG C, 120rpm shaken cultivation 16h is to optimal
Luminance, obtains test bacterium solution.
(5) impact that pH is grown to Vibrio-qinghaiensis sp. Q67：
The fluid medium that before sterilizing, step (3) is prepared by per bottle of 100mL subpackage to 7 250mL culture bottles, with
0.1mol/L NaOH solution and 0.1mol/L HCl solution adjust pH value respectively obtain pH be 6,6.5,7,7.5,8,8.5,9 liquid
Body culture medium, is inoculated with 100 μ L logarithmic (log) phase bacteria suspensions in superclean bench to 100mL fluid medium, 25 DEG C, 120rpm shakes
Culture 16h being swung to optimal luminescent state, tests its OD value and luminous intensity values respectively.Experimental result is as shown in figure 3, culture medium
PH has considerable influence to the growth of Vibrio-qinghaiensis sp. Q67, and Vibrio-qinghaiensis sp. Q67 upgrowth situation in the culture fluid that pH is 8 is strong with luminous
Degree is optimal, and for the culture medium that is prepared pH 7.95 or so, be not required to carry out the regulation of pH when therefore preparing culture medium.
(6) impact that temperature is grown to Vibrio-qinghaiensis sp. Q67：
The fluid medium that before sterilizing, step (3) is prepared by per bottle of 100mL subpackage to 5 250mL culture bottles, in
In superclean bench, 100 μ L logarithmic (log) phase bacteria suspensions of inoculation are respectively placed in 15,20,25,30,35 to 100mL fluid medium
DEG C, 120rpm shaken cultivation 16h to optimal luminescent state, test its OD value and luminous intensity values respectively.Experimental result such as Fig. 4 institute
Showing, when 25 DEG C, the upgrowth situation of Qinghai Vibrion is best, therefore selects bath temperature to be kept during shaken cultivation at 25 DEG C.
(7) bacterium solution luminous intensity is determined：
Negative high voltage being set as 800V, the bacterium solution 2mL standing 5min after culture in 16 hours is taken after superweak luminescence detector
Upper measure luminous value.Bacterium solution luminous intensity is 1.0 × 105Left and right can be used as test bacterium solution.
(8) flow injection system programming：Set intelligence flowing sample injection device program as shown in table 1.
(9) flow injection Luminous bacteria detection：
The test that step (4) is continuously cultivated passes through flow injection injector with bacterium solution, with the phosphate buffer that pH is 6.5
Superweak luminescence detecting system is pumped into as current-carrying, using ultra-pure water as blank, after obtaining stable luminous signal, successively
Pump into 3-nitropropionic acid solution, colchicine solution, Caulis Sacchari sinensis sample solution or the fresh Hemerocallis citrina Baroni of the variable concentrations that step (1) is prepared
Dish sample solution；Above-mentioned solution is first and test bacterium solution carries out premixing, then pumps into superweak luminescence detecting system through current-carrying
Carry out luminous detection and obtain corresponding luminous intensity.
(10) data integration and process and the design of warning system：Data integration and process and warning system system such as Fig. 7
Shown, by the photomultiplier tube being sequentially connected, opto-electronic conversion and voltage amplification module, analog-to-digital conversion module, detection by quantitative module
Constitute with alarm；The optical signal that step (9) detection is obtained is put by opto-electronic conversion voltage after photomultiplier tube amplification
Greatly, digital signal being obtained through analog digital conversion, is obtaining relative luminous intensity through detection by quantitative module.Relative luminous intensity=
(sample luminous intensity/control luminous intensity) × 100%, when detection by quantitative module detects relative luminous intensity less than 80%
Triggering alarm equipment alarm.
3-nitropropionic acid flow injection Luminous bacteria testing result is as shown in Figure 5：The minimum early warning of 3-nitropropionic acid is dense
Degree (relative luminous intensity is 80%) is 25.4mg/L.In rubescent Caulis Sacchari sinensis sample, the concentration of 3-nitropropionic acid is 36.3mg/L,
In early warning range.
Colchicine flow injection Luminous bacteria testing result is as shown in Figure 6：The minimum early warning concentration (phase of Colchicine
It is 80% to luminous intensity) it is 2.7mg/L, the Colchicine concentration for detecting in new hemerocallis sample is 3.4mg/L pre-
In alert scope.Therefore flow injection photobacteria detection method of the present invention can be used for the dirt of sudden food-borne toxin in drinking water
Dye.
Above-described embodiment is the present invention preferably embodiment, but embodiments of the present invention are not by above-described embodiment
Limit, other any spirit without departing from the present invention and the change that is made under principle, modification, replacement, combine, simplify,
Equivalent substitute mode is all should be, is included within protection scope of the present invention.
Claims (10)
1. the flow injection Luminous bacteria of a kind of quick detection and early warning food-borne endotoxin contamination, it is characterised in that include as
Lower step：
(1) recovery of Vibrio-qinghaiensis sp. Q67 microbial inoculum, inoculation and continuous culture；
(2) flow injection system programming；
(3) flow injection Luminous bacteria detection；
(4) data integration and process and the design of warning system.
2. the flow injection Luminous bacteria of quick detection according to claim 1 and early warning food-borne endotoxin contamination, its
It is characterised by, the comprising the following steps that of the flow injection Luminous bacteria：
(1) recovery of Vibrio-qinghaiensis sp. Q67 microbial inoculum, inoculation and continuous culture：- 20 DEG C of Vibrio-qinghaiensis sp. Q67 lyophilized powder ampoules 1 are taken, is disappeared
After poison, 4 DEG C of pre-cooling 15min, resuscitation fluid slight oscillatory is added, 1000 μ L bacteria suspensions is inoculated with superclean bench to 1000mL liquid
In body culture medium, 25 DEG C, 120rpm shaken cultivation 16h to optimal luminescent state, obtain testing with bacterium solution and being transferred to continuous training
Hair care fermentation tank is continuously cultivated；
(2) flow injection system programming：Flow injection injector program setting such as table 1；
Table 1 sets flow injection injector program
(3) flow injection Luminous bacteria detection：The test that step (1) is continuously cultivated passes through flow injection injector with bacterium solution
Superweak luminescence detecting system is pumped into, using ultra-pure water as blank, after obtaining stable luminous signal, imports testing sample
Solution carries out luminous detection and obtains corresponding luminous intensity；
(4) data integration and process and the design of warning system：Design data is integrated and processes and warning system is by being sequentially connected
Photomultiplier tube, opto-electronic conversion and voltage amplification module, analog-to-digital conversion module, detection by quantitative module and alarm composition；Step
Suddenly after the optical signal that (3) detection is obtained amplifies through photomultiplier tube, by opto-electronic conversion and voltage amplification module, then through mould
Number modular converter obtains digital signal, obtains relative luminous intensity through detection by quantitative module；Relative luminous intensity=(sample lights
Intensity/control luminous intensity) × 100%, the triggering when detection by quantitative module detects relative luminous intensity less than 80% is reported to the police
Device is reported to the police.
3. the flow injection Luminous bacteria of quick detection according to claim 1 and early warning food-borne endotoxin contamination, its
It is characterised by, step (1) fluid medium is prepared according to the following steps and is transferred to feed supplement fluid reservoir after carrying out sterilization treatment
In：Weigh tryptone 5g, yeast powder 5g, NaCl 6.73g, Na2CO30.99g、NaHCO30.57g、MgCl2·6H2O
2.92g、MgSO42.47g、CaCO30.03g, NaBr 0.1g and KCl 0.22g, is added in the conical flask of 1L one by one,
Glycerol adding 3mL, plus distilled water 1000mL, mixed elimination precipitation, with the rear 121 DEG C of sterilizings 15min of kraft paper wrapping.
4. the flow injection Luminous bacteria of quick detection according to claim 1 and early warning food-borne endotoxin contamination, its
It is characterised by, step (3) sample solution is the aqueous solution containing 3-nitropropionic acid or containing Colchicine.
5. the flow injection Luminous bacteria of quick detection according to claim 1 and early warning food-borne endotoxin contamination, its
It is characterised by, step (3) sample solution is drinking water, the water source of food processing enterprises, Caulis Sacchari sinensis sample solution or new cadmium yellow
Herba Astragali Sinici sample solution.
6. the flow injection Luminous bacteria of quick detection according to claim 5 and early warning food-borne endotoxin contamination, its
Be characterised by, step (3) testing sample solution be drinking water when, in step (4) relative luminous intensity below 80% then
Show that drinking water is potentially contaminated, alarm equipment alarm is triggered after detection by quantitative module analysis.
7. the flow injection Luminous bacteria of quick detection according to claim 1 and early warning food-borne endotoxin contamination, its
It is characterised by, in step (3) detection process, test bacterium solution and sample solution shift to an earlier date premixing and then by phosphate buffer
It is loaded into superweak luminescence detecting system to be detected.
8. the flow injection Luminous bacteria of quick detection according to claim 7 and early warning food-borne endotoxin contamination, its
It is characterised by, incorporation time in detection process is 10s to step (3) sample solution with test bacterium solution, and detection time is
60s.
9. the flow injection Luminous bacteria of quick detection according to claim 1 and early warning food-borne endotoxin contamination, its
It is characterised by, in step (3) detection process, test bacterium solution feed rate is adjusted according to detection speed to ensure the bacterium solution beginning
The luminous intensity of relative constancy is kept eventually.
10. the flow injection Luminous bacteria of quick detection as claimed in claim 1 and early warning food-borne endotoxin contamination is in food
Sudden food-borne endotoxin contamination detection in the pollution of waterhead monitoring of product processing enterprise, drinking water on-line monitoring and drinking water
In application.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611041464.2A CN106442483A (en) | 2016-11-22 | 2016-11-22 | Luminous bacterium flow injection method for quickly detecting and warning food-borne toxin pollution and application of luminous bacterium flow injection method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611041464.2A CN106442483A (en) | 2016-11-22 | 2016-11-22 | Luminous bacterium flow injection method for quickly detecting and warning food-borne toxin pollution and application of luminous bacterium flow injection method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106442483A true CN106442483A (en) | 2017-02-22 |
Family
ID=58218024
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611041464.2A Pending CN106442483A (en) | 2016-11-22 | 2016-11-22 | Luminous bacterium flow injection method for quickly detecting and warning food-borne toxin pollution and application of luminous bacterium flow injection method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106442483A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109828057A (en) * | 2019-03-27 | 2019-05-31 | 湖北省食品质量安全监督检验研究院 | A kind of sugarcane neurotoxin efficiently extracts and its liquid chromatography detecting method |
| CN111315860A (en) * | 2017-11-10 | 2020-06-19 | 横河电机株式会社 | Microorganism contamination countermeasure selection device, microorganism contamination countermeasure selection system, microorganism contamination countermeasure selection method, and microorganism contamination countermeasure selection program |
| CN113340885A (en) * | 2021-06-23 | 2021-09-03 | 同济大学 | Formaldehyde biological monitoring device and monitoring method |
| CN113438955A (en) * | 2019-01-25 | 2021-09-24 | 韦务拓客公司 | Human body injectate delivery device and method and catheter device |
| CN114813592A (en) * | 2022-03-11 | 2022-07-29 | 华南理工大学 | Method for researching degradation mechanism of hydroxychloroquine in natural water body |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1343888A (en) * | 2001-10-19 | 2002-04-10 | 厦门大学 | Photogen sensor device for monitoring water toxin |
| WO2006125954A2 (en) * | 2005-05-26 | 2006-11-30 | Cymtox Limited | Water monitoring system |
| CN101871927A (en) * | 2010-06-13 | 2010-10-27 | 南京大学 | Online monitoring equipment and method for acute toxicity of water quality |
| CN105987898A (en) * | 2015-03-05 | 2016-10-05 | 上海市南洋模范中学 | Detection method for acute biological toxicities of pollutants on dust-haze day |
-
2016
- 2016-11-22 CN CN201611041464.2A patent/CN106442483A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1343888A (en) * | 2001-10-19 | 2002-04-10 | 厦门大学 | Photogen sensor device for monitoring water toxin |
| WO2006125954A2 (en) * | 2005-05-26 | 2006-11-30 | Cymtox Limited | Water monitoring system |
| CN101218499A (en) * | 2005-05-26 | 2008-07-09 | 希姆陶克斯有限公司 | Water monitoring system |
| CN101871927A (en) * | 2010-06-13 | 2010-10-27 | 南京大学 | Online monitoring equipment and method for acute toxicity of water quality |
| CN105987898A (en) * | 2015-03-05 | 2016-10-05 | 上海市南洋模范中学 | Detection method for acute biological toxicities of pollutants on dust-haze day |
Non-Patent Citations (1)
| Title |
|---|
| 周姣: "流动注射法监测食品用水中重金属突发性污染", 《中国优秀硕士学位论文全文数据库 工程科技Ⅰ辑》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111315860A (en) * | 2017-11-10 | 2020-06-19 | 横河电机株式会社 | Microorganism contamination countermeasure selection device, microorganism contamination countermeasure selection system, microorganism contamination countermeasure selection method, and microorganism contamination countermeasure selection program |
| CN113438955A (en) * | 2019-01-25 | 2021-09-24 | 韦务拓客公司 | Human body injectate delivery device and method and catheter device |
| CN109828057A (en) * | 2019-03-27 | 2019-05-31 | 湖北省食品质量安全监督检验研究院 | A kind of sugarcane neurotoxin efficiently extracts and its liquid chromatography detecting method |
| CN113340885A (en) * | 2021-06-23 | 2021-09-03 | 同济大学 | Formaldehyde biological monitoring device and monitoring method |
| CN114813592A (en) * | 2022-03-11 | 2022-07-29 | 华南理工大学 | Method for researching degradation mechanism of hydroxychloroquine in natural water body |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106442483A (en) | Luminous bacterium flow injection method for quickly detecting and warning food-borne toxin pollution and application of luminous bacterium flow injection method | |
| AU660048B2 (en) | Apparatus for monitoring liquids | |
| Del Giorgio et al. | Coherent patterns in bacterial growth, growth efficiency, and leucine metabolism along a northeastern Pacific inshore‐offshore transect | |
| ITRM20120218A1 (en) | DEVICE AND METHOD FOR ANALYSIS AND MONITORING OF TOXICITY IN WATERS. | |
| CN106755286B (en) | A method of testing oil extraction waste water bio-toxicity using Vibrio-qinghaiensis sp. Q67 | |
| US20040171094A1 (en) | Oxygen sensors disposed on a microtiter plate | |
| CN102495051A (en) | Device and method for quickly detecting biological activity and metabolism | |
| CN102288653A (en) | Online biochemical oxygen demand (BOD) detector and detection method of same | |
| CN108107025A (en) | A kind of water quality detection method and system | |
| CN103336045A (en) | Device for on-line detection and automatic alarm of toxic substances and detection method of toxic substances | |
| CN106018688B (en) | A kind of evaluation method of metal nanoparticle ion and nano effect toxicity contribution rate | |
| Welschmeyer et al. | A portable, sensitive plankton viability assay for IMO shipboard ballast water compliance testing | |
| CN105203510B (en) | A kind of microorganism in food rapid detection method | |
| CN202083662U (en) | Online tester for biochemical oxygen demand BOD | |
| CN109781682B (en) | Method for determining soluble organic matters in breeding wastewater duckweed treatment system | |
| Garzón-Cardona et al. | Chemically unidentified dissolved organic carbon: A pivotal piece for microbial activity in a productive area of the Northern Patagonian shelf | |
| CN109884015A (en) | The method of application and detection CHL of the MOF-Zn fluorescent optical sensor in detection chloramphenicol | |
| CN102759517B (en) | Analytical instrument and method for evaluating microbial contamination of an object | |
| CN109580514B (en) | A method of measuring abiotic particle cluster flow and biologic grain state phosphorus content | |
| CN201006428Y (en) | Sulphate reduction bacterium testing bottle | |
| CN208187983U (en) | A kind of blood culture report sun detection device | |
| CN103940789B (en) | The specification of photobacteria Fluorescence behaviour and numeric value analysis in water toxicity detection | |
| CN111394348A (en) | Method for extracting and detecting free DNA in sewage | |
| CN103018419B (en) | Method for determining risk of toxic pollutants in river water of high content of sand | |
| CN1584554A (en) | Photochemical and biochemical microorganic membrane dynamic responding sensor for COD with optical fiber |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20170222 |